Accuracy of replication in the polymerase chain reaction. Comparison between Thermotoga maritima DNA polymerase and Thermus aquaticus DNA polymerase

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dc.contributor.authorDiaz, Ricardo Sobhie [UNIFESP]
dc.contributor.authorSabino, Ester Cerdeira [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.contributor.institutionIrwin Memorial Blood Centers
dc.contributor.institutionFundação Pró-Sangue, Hemocentro de São Paulo
dc.date.accessioned2015-06-14T13:24:47Z
dc.date.available2015-06-14T13:24:47Z
dc.date.issued1998-10-01
dc.description.abstractFor certain applications of the polymerase chain reaction (PCR), it may be necessary to consider the accuracy of replication. The breakthrough that made PCR user friendly was the commercialization of Thermus aquaticus (Taq) DNA polymerase, an enzyme that would survive the high temperatures needed for DNA denaturation. The development of enzymes with an inherent 3' to 5' exonuclease proofreading activity, lacking in Taq polymerase, would be an improvement when higher fidelity is needed. We used the forward mutation assay to compare the fidelity of Taq polymerase and Thermotoga maritima (ULTMA ) DNA polymerase, an enzyme that does have proofreading activity. We did not find significant differences in the fidelity of either enzyme, even when using optimal buffer conditions, thermal cycling parameters, and number of cycles (0.2% and 0.13% error rates for ULTMA and Taq, respectively, after reading about 3,000 bases each). We conclude that for sequencing purposes there is no difference in using a DNA polymerase that contains an inherent 3' to 5' exonuclease activity for DNA amplification. Perhaps the specificity and fidelity of PCR are complex issues influenced by the nature of the target sequence, as well as by each PCR component.en
dc.description.affiliationUniversidade Federal de São Paulo (UNIFESP)
dc.description.affiliationIrwin Memorial Blood Centers
dc.description.affiliationFundação Pró-Sangue, Hemocentro de São Paulo
dc.description.affiliationUnifespUNIFESP, EPM, São Paulo
dc.description.sourceSciELO
dc.format.extent1239-1242
dc.identifierhttp://dx.doi.org/10.1590/S0100-879X1998001000001
dc.identifier.citationBrazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 31, n. 10, p. 1239-1242, 1998.
dc.identifier.doi10.1590/S0100-879X1998001000001
dc.identifier.fileS0100-879X1998001000001.pdf
dc.identifier.issn0100-879X
dc.identifier.scieloS0100-879X1998001000001
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/679
dc.identifier.wosWOS:000076614900001
dc.language.isoeng
dc.publisherAssociação Brasileira de Divulgação Científica
dc.relation.ispartofBrazilian Journal of Medical and Biological Research
dc.rightsAcesso aberto
dc.subjectpolymerase chain reaction fidelityen
dc.subjectThermus aquaticus (Taq) DNA polymeraseen
dc.subjectThermotoga maritima DNA polymeraseen
dc.titleAccuracy of replication in the polymerase chain reaction. Comparison between Thermotoga maritima DNA polymerase and Thermus aquaticus DNA polymeraseen
dc.typeArtigo
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